Pheochromocytoma-related cardiomyopathy presenting as acute myocardial infarction: A case report.
Pheochromocytoma (PHEO)-related cardiomyopathy is a rare condition in which release of a large amount of catecholamines leads to severe vasoconstriction, coronary vasospasm, myocardial ischemia, injury, and necrosis. Its clinical manifestations can be similar to those of acute coronary syndrome. A 63-year-old woman was diagnosed with acute non-ST segment elevation myocardial infarction following chest pain for 8 hours. The results of coronary angiography were normal. The patient developed dyspnea, cough with frothy pink sputum, paroxysmal sweating, arrhythmia, and blood pressure fluctuation, and was transferred to the intensive care unit for monitoring and treatment. PHEO, catecholamine cardiomyopathy (CICMP). After monitoring the pulse index continuous cardiac output and treatment with α and β adrenergic receptor blockers for 18 days, laparoscopic resection of the left adrenal mass was performed. The patient's condition improved and she was discharged 31 days after admission. Outpatient follow-up examinations 1 month and 1 year later did not show recurrence. PHEO can cause CICMP, the manifestations of which are partly similar to those of takotsubo cardiomyopathy (TTC). Once the patient's condition stabilizes, surgery should be considered. Fluid management is necessary, and agents such as α and β adrenergic receptor blockers should be administered.


MIBG superscan of metastatic paraganglioma occurring with neurofibromatosis type 1.
A Tc MDP bone superscan occurs when osseous activity is extremely intense and genitourinary and soft tissue activity is not identified. A similar phenomenon has been described with metaiodobenzylguanidine (MIBG) in metastatic pheochromocytoma and neuroblastoma. We present a case of metastatic paraganglioma resulting in an MIBG superscan. Neuroendocrine bone metastasis alters the biodistribution of MIBG such that the liver, heart, and urinary bladder are not well visualized. Our case occurred in association with neurofibromatosis type 1 and in the absence of an identified primary tumor.


Attenuation of an original US porcine epidemic diarrhea virus strain PC22A via serial cell culture passage.
Although porcine epidemic diarrhea (PED) has caused huge economic losses in the pork industry worldwide, an effective live, attenuated vaccine is lacking. In this study, an original US, highly virulent PED virus (PEDV) strain PC22A was serially passaged in Vero CCL81 and Vero BI cells. The virus growth kinetics in cell culture, virulence in neonatal pigs and the whole genomic sequences of selected passages were examined. Increased virus titers and sizes of syncytia were observed at the 65th passage level (P65) and P120, respectively. Based on the severity of clinical signs, histopathological lesions and the distribution of PEDV antigens in the gut, the virulence of P100 and above, but not P95C13 (CCL81), was markedly reduced in 4-day-old, caesarian-derived, colostrum-deprived piglets. Subsequently, the attenuation of P120 and P160 was confirmed in 4-day-old, conventional suckling piglets. Compared with P120, P160 replicated less efficiently in the intestine of pigs and induced a lower rate of protection after challenge. Sequence analysis revealed that the virulent viruses [P3 and P95C13 (CCL81)] had one, one, sixteen (including an early termination of nine amino acids) and two amino acid differences in non-structure protein 1 (nsp1), nsp4, spike and membrane proteins, respectively, from the fully attenuated P160. However, the overall pattern of attenuation-related genetic changes in PC22A differed from those of the other four pairs of PEDV wild type strains and their attenuated derivatives. These results suggest that PEDV attenuation can occur through multiple molecular mechanisms. The knowledge provides insights into potential molecular mechanisms of PEDV attenuation.


The use of fish and herptiles in traditional folk therapies in three districts of Chenab riverine area in Punjab, Pakistan.
Like botanical taxa, various species of animals are also used in traditional and modern health care systems. Present study was intended with the aim to document the traditional uses of herptile and fish species among the local communities in the vicinity of the River Chenab, Punjab Pakistan. Data collected by semi-structured interviews and questionnaires were subsequently analyzed using relative frequency of citation (FC), fidelity level (FL), relative popularity level (RPL), similarity index (SI), and rank order priority (ROP) indices. Out of total 81 reported species, ethnomedicinal uses of eight herptiles viz. Aspideretes gangeticus, A. hurum, Eublepharis macularius, Varanus bengalensis, Python molurus, Eryx johnii, Ptyas mucosus mucosus, Daboia russelii russelii and five fish species including Hypophthalmichthys molitrix, Cirrhinus reba, Labeo dero, Mastacembelus armatus, and Pethia ticto were reported for the first time from this region. Fat, flesh, brain, and skin were among the commonly utilized body parts to treat allergy, cardiovascular, nervous and respiratory disorders, sexual impotency, skin infections, and as antidote and anti-diabetic agents. Hoplobatrachus tigerinus, Duttaphrynus stomaticus, and Ptyas mucosus mucosus (herptiles), as well as Labeo rohita, Wallago attu, and Cirrhinus reba (fish) were top ranked with maximum informant reports, frequency of citations, and rank order priority. Uromastyx hardwickii, Ctenopharyngodon idella, H. molitrix, Cirrhinus mrigala, C. reba, L. rohita, L. calbasu, L. dero, and Pethia ticto were the species with 100% fidelity level. Furthermore, medicinal uses of Aspideretes gangeticus, Aspideretes hurum, Calotes versicolor, Daboia russelii russelii, Hypophthalmichthys molitrix, Cirrhinus reba, Labeo dero, Mastacembelus armatus, Pethia ticto, and Gagata cenia were reported for the first time. About half of the reported species depicted zero similarity index with previously reported literature, which indicates strong associations of local inhabitants with animal species, particularly for therapeutic purpose. Inclusive studies on composition and bioactivities of the species with maximum use reports may contribute significantly in animal-based novel drugs discovery.


[The characteristics of the antibacterial activity of garlic (author's transl)].
It has been found by Cavallito (1944) that garlic (Allium sativum L.) has a strong antibacterial activity and the active principle of garlic is named "Allicin" and it has been confirmed by Stoll et al. (1948) that the allicin is derived from the alliin-alliinase system. Nevertheless, one worker in Japan (1951) stands on another view point which the effective component of garlic is some sugar-proteim because he could not find alliin in the native garlic. We intend to make sure that the alliin is contained in the native garlic, by using Hitachi Amino acid Analyser with Fraction Collecter and compared with garlic of other countrys' production. As the result of examinations, we found that the good amounts of alliin is contained in the native garlic also. The contents of alliin in garlic is varied by the species, the cultivation mode, and the district of production. The form of alliin in the tissue is a free style, and thereupon it is easy to change its chemical conformation by the method of extraction. We compared the relative capacity between 1 allicin unit by Klein (1954) and 1 cup unit by Schmidt et al. (1947). Furthermore we tested the antibiotic spectrum, using the pure crystallized alliin with the sufficient amount of raw alliinase gained by the isoelectric point precipitation and checked upon wide spectrum for Mycobacterium tuberculosis, Gram-positive and negative bacteria, and some fungi.


Temperature dependent electrical properties of organic light-emitting diodes using a Zn complex.
We synthesized a new electroluminescent material [(1,10-phenanthroline)(8-hydroxyqu inoline)] Zn(phen)q. The temperature dependence of the electrical properties of OLED using Zn(phen)q are studied to understand the electrical conduction mechanism. The current density-voltage characteristics are measured in the temperature range of 60 approximately 240 K and analyzed using a hopping model with an exponential trap distribution. At temperatures above 150 K, hopping conduction with an exponential trap distribution is dominant. We have obtained a characteristic trap depth of E(t) = 0.13 eV.


Hydrogen deuterium exchange defines catalytically linked regions of protein flexibility in the catechol O-methyltransferase reaction.
Human catechol O-methyltransferase (COMT) has emerged as a model for understanding enzyme-catalyzed methyl transfer from S-adenosylmethionine (AdoMet) to small-molecule catecholate acceptors. Mutation of a single residue (tyrosine 68) behind the methyl-bearing sulfonium of AdoMet was previously shown to impair COMT activity by interfering with methyl donor-acceptor compaction within the activated ground state of the wild type enzyme [J. Zhang, H. J. Kulik, T. J. Martinez, J. P. Klinman, Proc. Natl. Acad. Sci. U.S.A. 112, 7954-7959 (2015)]. This predicts the involvement of spatially defined protein dynamical effects that further tune the donor/acceptor distance and geometry as well as the electrostatics of the reactants. Here, we present a hydrogen/deuterium exchange (HDX)-mass spectrometric study of wild type and mutant COMT, comparing temperature dependences of HDX against corresponding kinetic and cofactor binding parameters. The data show that the impaired Tyr68Ala mutant displays similar breaks in Arrhenius plots of both kinetic and HDX properties that are absent in the wild type enzyme. The spatial resolution of HDX below a break point of 15-20 °C indicates changes in flexibility across ∼40% of the protein structure that is confined primarily to the periphery of the AdoMet binding site. Above 20 °C, Tyr68Ala behaves more like WT in HDX, but its rate and enthalpic barrier remain significantly altered. The impairment of catalysis by Tyr68Ala can be understood in the context of a mutationally induced alteration in protein motions that becomes manifest along and perpendicular to the primary group transfer coordinate.


Dispersal and population genetic structure of Telmatherina antoniae, an endemic freshwater Sailfin silverside from Sulawesi, Indonesia.
Population genetic structure in the presence of substantial dispersal provides a unique perspective on the evolution of reproductive isolation. We sampled Telmatherina antoniae, an endemic fish species, at 10 sites in Lake Matano, Indonesia. Significant genetic structure (F(ST) = 0.03) was found, despite a migration rate of 10.2% and a mean dispersal distance of 13.6 km, estimated by genotype assignment. Neither dispersal distance nor direction differed from random expectations, indicative of no dispersal barrier in Lake Matano. However, Bayesian genotype cluster assignment identified a population structure consisting of four to six clusters that did not coincide with sample site distribution, but explained two to three times more genetic variance than sample site. The mechanism for continued isolation of those genetic clusters is unknown, but assortative mating and temporal isolation are obvious candidates. Our results resolve the apparent paradox of population genetic structure coupled with frequent dispersal, and highlight the importance of considering cryptic genetic structure.


Molecular recognition of paxillin LD motifs by the focal adhesion targeting domain.
Focal adhesions (FAs) are large submembrane signaling complexes formed at sites of cellular attachment to the extracellular matrix. The interaction of LD motifs with their targets plays an important role in the assembly of FAs. We have determined the molecular basis for the recognition of two paxillin LD motifs by the FA targeting (FAT) domain of FA kinase using a combination of X-ray crystallography, solution NMR, and homology modeling. The four-helix FAT domain displays two LD binding sites on opposite sites of the molecule that bind LD peptides in a helical conformation. Threading studies suggest that the LD-interacting domain of p95PKL shares a common four-helical core with the FAT domain and the tail of vinculin, defining a structural family of LD motif binding modules.


CONSTRICTOR: constraint modification provides insight into design of biochemical networks.
Advances in computational methods that allow for exploration of the combinatorial mutation space are needed to realize the potential of synthetic biology based strain engineering efforts. Here, we present Constrictor, a computational framework that uses flux balance analysis (FBA) to analyze inhibitory effects of genetic mutations on the performance of biochemical networks. Constrictor identifies engineering interventions by classifying the reactions in the metabolic model depending on the extent to which their flux must be decreased to achieve the overproduction target. The optimal inhibition of various reaction pathways is determined by restricting the flux through targeted reactions below the steady state levels of a baseline strain. Constrictor generates unique in silico strains, each representing an "expression state", or a combination of gene expression levels required to achieve the overproduction target. The Constrictor framework is demonstrated by studying overproduction of ethylene in Escherichia coli network models iAF1260 and iJO1366 through the addition of the heterologous ethylene-forming enzyme from Pseudomonas syringae. Targeting individual reactions as well as combinations of reactions reveals in silico mutants that are predicted to have as high as 25% greater theoretical ethylene yields than the baseline strain during simulated exponential growth. Altering the degree of restriction reveals a large distribution of ethylene yields, while analysis of the expression states that return lower yields provides insight into system bottlenecks. Finally, we demonstrate the ability of Constrictor to scan networks and provide targets for a range of possible products. Constrictor is an adaptable technique that can be used to generate and analyze disparate populations of in silico mutants, select gene expression levels and provide non-intuitive strategies for metabolic engineering.


P27 expression in normal epithelium and condylomas of the vulva in HIV positive and negative women.
The objective of the present study was to evaluate the expression of the p27 protein in the normal epithelium and vulvar condylomas in human immunodeficiency (HIV) positive and negative patients. Eight samples of normal vulvar epithelium were evaluated (Group A), ten of the HIV negative vulvar condyloma patients (Group B) and another eight of the vulvar condyloma HIV positive patients (Group C). The DNA of human papillomavirus (HPV) was identified by means of polymerase chain reaction (PCR). Immunohistochemistry was the method used to evaluate the expression of p27 using monoclonal mouse antibody (Monoclonal Mouse, anti-human p27, Clone Sx 53 G8). The immunoexpression was evaluated at a magnification of 400x, counting a minimum of 1,000 cells per slide. The results obtained were the following: a) comparing groups A and B and groups A and C there was a significant difference in relation to the expression of the p27 protein which was 63.32% in group A and only 13.35% and 18.89% in groups B and C, respectively; b) comparing groups B and C among them, there was no significant difference. We concluded that in normal vulvar tissue the p27 protein is present in a large number of cells and that in vulval condylomas its expression is very much lowered both in HIV positive and negative cases.


Microstructure, Magnetic, and Magnetoresistance Properties of La0.7Sr0.3MnO3:CuO Nanocomposite Thin Films.
(La0.7Sr0.3MnO3)0.67:(CuO)0.33 (LSMO:CuO) nanocomposite thin films were deposited on SrTiO3 (001), LaAlO3 (001), and MgO (001) substrates by pulsed laser deposition, and their microstructure as well as magnetic and magnetoresistance properties were investigated. X-ray diffraction (XRD) and transmission electron microscopy (TEM) results show that LSMO:CuO films grow as highly textured self-assembled vertically aligned nanocomposite (VAN), with a systematic domain structure and strain tuning effect based on the substrate type and laser deposition frequency. A record high low-field magnetoresistance (LFMR) value of ∼80% has been achieved in LSMO:CuO grown on LaAlO3 (001) substrate under high frequency. Detailed analysis indicates that both the strain state and the phase boundary effect play a significant role in governing the overall LFMR behavior.


IP3-independent signalling of OX1 orexin/hypocretin receptors to Ca2+ influx and ERK.
OX1 orexin receptors (OX1R) have been shown to activate receptor-operated Ca2+ influx pathways as their primary signalling pathway; however, investigations are hampered by the fact that orexin receptors also couple to phospholipase C, and therewith inositol-1,4,5-trisphosphate (IP3)-dependent Ca2+ release. We have here devised a method to block the latter signalling in order to focus on the mechanism of Ca2+ influx activation by OX1R in recombinant systems. Transient expression of the IP3-metabolising enzymes IP3-3-kinase-A (inositol-1,4,5-trisphosphate-->inositol-1,3,4,5-tetrakisphosphate) and type I IP3-5-phosphatase (inositol-1,4,5-trisphosphate-->inositol-1,4-bisphosphate) almost completely attenuated the OX1R-stimulated IP3 elevation and Ca2+ release from intracellular stores. Upon attenuation of the IP3-dependent signalling, the receptor-operated Ca2+ influx pathway became the only source for Ca2+ elevation, enabling mechanistic studies on the receptor-channel coupling. Attenuation of the IP3 elevation did not affect the OX1R-mediated ERK (extracellular signal-regulated kinase) activation in CHO cells, which supports our previous finding of the major importance of receptor-operated Ca2+ influx for this response.


Effective communication with prescribers: tips and insights from both sides of the fence.
Although effective collaboration between pharmacists and prescribers is critically important, their relations are often strained. Lack of or inadequate communication of the medication-related problem can hinder pharmacists from receiving timely responses to their recommendations and affect the quality of care provided to the patient. Here, seasoned consultant pharmacists and medical directors suggest simple strategies to help cultivate more collaborative, effective, and satisfying relationships.


Spinal cord injury elicits expression of keratan sulfate proteoglycans by macrophages, reactive microglia, and oligodendrocyte progenitors.
Keratan sulfate proteoglycans (KSPGs) are extracellular matrix molecules that appear to establish boundaries for axonal growth in the developing brain and spinal cord. In vitro studies confirm that KSPGs define inhibitory boundaries to extending neurites. The aim of the current study was to investigate whether KSPGs are expressed after spinal cord injury (SCI) and thereby might act as potential inhibitors of axonal growth. Adult Fischer 344 rats were subjected to spinal cord lesions, and the temporal and spatial expression of KSPGs was examined using the 5D4 monoclonal anti-KSPG antibody. In the intact spinal cord, a subpopulation of microglia expressed 5D4-KSPG throughout the white and gray matter. Within 24 hr of injury, 5D4-KSPG immunoreactivity substantially increased and appeared on cellular profiles in close proximity to the spinal cord lesion site, peaking 3 d after injury. Double immunolabeling revealed that 5D4-KSPG expression arose from multiple cell types at the lesion site, including reactive microglia, macrophages, and oligodendrocyte progenitors. Astrocytes were not identified as a source of 5D4-KSPG. The robust and extensive production of 5D4-KSPG at sites of SCI precedes the expression of other putatively inhibitory proteoglycan molecules such as chondroitin sulfate proteoglycans. This is the first demonstration that KSPGs are expressed after SCI in a temporal and spatial relationship that could exert an early and important role in modulating axonal growth after SCI.


Possible involvement of zinc deficiency in epidermal growth factor receptor inhibitor-induced xerotic dermatitis.
Cancer treatment with epidermal growth factor receptor inhibitors (EGFRIs) often induces severe xerotic dermatitis. Various irritants facilitate development of dermatitis in xerotic skin. As zinc deficiency plays a role in the development of irritant dermatitis, we measured serum zinc levels in 25 patients with xerotic dermatitis due to treatment with EGFRIs. Of these patients, nine were treated with EGFR tyrosine kinase inhibitors and 16 were treated with anti-EGFR antibody alone or in combination with other anticancer agents. Serum zinc levels of all patients were lower than the normal range of >80 μg/dL, with a mean ± SD serum zinc level of 56.4 ± 11.7 μg/dL. These were correlated with serum magnesium levels in patients. As the serum magnesium level is known to be reduced by the inhibition of EGFR, a similar mechanism may also be involved in decreasing the serum zinc level. Among 21 patients treated with zinc supplementation for more than 2 months, xerotic dermatitis markedly improved, with an increase of serum zinc levels in 16 patients. The other five patients exhibited no significant improvement in their skin condition, and insufficient and unstable increase in serum zinc levels. In conclusion, zinc supplementation may be beneficial in supportive care for patients with EGFRI-induced xerotic dermatitis.


Pathophysiology of stroke rehabilitation: the natural course of clinical recovery, use-dependent plasticity and rehabilitative outcome.
Even though the disruption of motor activity and function caused by stroke is at times severe, recovery is often highly dynamic. Recuperation reflects the ability of the neuronal network to adapt. Next to an unmasking of latent network representations, other adaptive processes, such as excitatory metabolic stress, an imbalance in activating and inhibiting transmission, leading to salient hyperexcitability, or the consolidation of novel connections, prime the plastic capabilities of the system. Rehabilitative interventions may modulate mechanisms of neurofunctional plasticity and influence the natural course after stroke, both positively, but potentially also acting detrimentally. Though routine rehabilitative procedures are an integral part of stroke care, evidence as to their effectiveness remains equivocal. The present review describes the natural course of motor recovery, focusing on ischemic stroke, and discusses use- and training-dependent adaptive effects. It complements a prior article which highlighted the pathophysiology of plasticity. Though the interaction between rehabilitation and plasticity remains elusive, an attempt is made to clarify how and to what extent rehabilitative therapy shapes motor recovery.


Parental perceptions of barriers to mental health services for young people.
This study explores a range of barriers that parents encountered in accessing mental health services. The study also explored whether parents experienced similar barriers to accessing services in 2003 and 2013. One hundred and thirty-four parents of young people attending an initial assessment at a Child and Adolescent Mental Health Service (CAMHS) or headspace centre completed a questionnaire assessing 10 general barriers to care. These data were compared to those collected from 129 participants at CAMHS in 2003. The ranking of barriers to mental health care for their children was similar for both survey years, with 'wait time being too long' and 'help being too expensive' the two highest ranked barriers. Cost factors were related to not knowing that the services did not charge fees and having to take time off work to attend appointments. Multiple referral steps and uncertain wait times were the main concerns regarding wait times. The overall strength with which barriers were endorsed remained relatively low; however, at least 40% of the sample agreed they had experienced four of the barriers in both years. Despite relatively low endorsement of barriers, there are substantial proportions of parents who experienced some barriers to services, and services should continue working to reduce them to facilitate timely access. There is a particular need for more service-related information to clarify that public sector mental health services do not charge fees. Methods such as rapid initial assessment and actively managing wait lists may go some way to reducing perceived wait time barriers.


Urban air quality forecasting based on multi-dimensional collaborative Support Vector Regression (SVR): A case study of Beijing-Tianjin-Shijiazhuang.
Today, China is facing a very serious issue of Air Pollution due to its dreadful impact on the human health as well as the environment. The urban cities in China are the most affected due to their rapid industrial and economic growth. Therefore, it is of extreme importance to come up with new, better and more reliable forecasting models to accurately predict the air quality. This paper selected Beijing, Tianjin and Shijiazhuang as three cities from the Jingjinji Region for the study to come up with a new model of collaborative forecasting using Support Vector Regression (SVR) for Urban Air Quality Index (AQI) prediction in China. The present study is aimed to improve the forecasting results by minimizing the prediction error of present machine learning algorithms by taking into account multiple city multi-dimensional air quality information and weather conditions as input. The results show that there is a decrease in MAPE in case of multiple city multi-dimensional regression when there is a strong interaction and correlation of the air quality characteristic attributes with AQI. Also, the geographical location is found to play a significant role in Beijing, Tianjin and Shijiazhuang AQI prediction.


Polymer-templated nanospider TiO(2) thin films for efficient photoelectrochemical water splitting.
We have discovered a facile and inexpensive approach to fabricate "nanospider" TiO(2) thin films with not only an amazing morphology but highly efficient water splitting to produce hydrogen. Our method employs benzene-swollen poly(ethylene glycol) as a sacrificial organic polymer to template the semiconductor thin film. The synthesized TiO(2) thin films are highly crystalline with optimized particle and channel size to enhance the liquid-semiconductor junction interaction. This enhanced contact area leads to more than twice the water splitting performance than conventional P25 thin films. In addition, the nanospider thin films also outperform P25 films in the photodegradation of toxic organics.


Computational Design of Oligopeptide Containing Poly(ethylene glycol) Brushes for Stimuli-Responsive Drug Delivery.
Stimuli-responsive biomaterials are used to facilitate drug and gene delivery by shielding the drug/gene during circulation times and selectively releasing the cargo at the desired target. Within stimuli-responsive materials, pH-responsive materials are exploited for delivery to specific organs, intracellular compartments, cancer cells, site of inflammation or infection as those sites are characterized by pH that is different from the blood pH. In this paper we use molecular dynamics (MD) simulations to design such pH-responsive biomaterials where the balance between the various intermolecular interactions (e.g., electrostatics, van der Waals) within the biomaterials allow biofunctional molecules to be reversibly shielded and exposed to the environment with change in pH. In our model the shielding aspect is imparted by a polyethylene glycol (PEG) brush and the pH-responsive component is a PEG-tethered oligopeptide that undergoes changes in conformations via protonation of residues upon changes in pH. Starting with a PEG-tethered peptide in a monodisperse short PEG brush, we first vary the composition and sequence of histidine (H), lysine (K), and glutamate (E) along the oligopeptide sequence to find the design parameters that maximize the shielding and exposure of the oligopeptide at pH ∼ 7.0 and pH < 7.0, respectively. Then, we probe the effect of the PEG brush on the conformations of the oligopeptides by simulating PEG-tethered peptide in a bimodal PEG brush containing short PEG and long PEG chains. We characterize the intermolecular interactions involving the PEG, peptide, and solvent that influence the shielded and exposed conformations of the oligopeptides at the two different pHs. In a short monodisperse PEG brush, with a longer PEG-tethered peptide containing large blocks of histidines that undergo change in protonation state as a response to pH change, placed between a protonated lysine and deprotonated glutamate, the PEG brush exhibits maximum shielding and exposure with pH change. This change from shielded to exposed state is driven by electrostatic repulsion upon H protonation. The presence of long PEG chains in a bimodal PEG brush leads to dominating PEG-peptide attractive interactions that reduces the contrast in shielded and exposed conformations of the PEG-tethered peptide upon protonation of histidines.


Coagulase-negative staphylococcal infections in a neonatal intensive care unit: In vivo response to cloxacillin.
Following the introduction of cloxacillin and gentamicin as the first line of treatment for possible late-onset sepsis (LOS) in the authors' neonatal intensive care unit (NICU), it was subsequently noted that very low birth weight (VLBW) infants improved clinically, despite subsequently positive blood cultures for oxacillin-resistant, coagulase-negative Staphylococcus (CONS). The results of the management of VLBW infants with CONS sepsis during one calendar year, based on clinical rather than laboratory findings, are presented. VLBW infants with LOS were identified through the neonatal database, and the charts of those with CONS were reviewed for antibiotic usage, antibiotic resistance pattern, clearance of CONS from the blood and NICU discharge status. Oxacillin sensitivity was determined by the presence of the mecA gene. From January 1 to December 31, 2002, 27 VLBW infants, treated in the authors' NICU for LOS due to CONS, were identified. The mean age of LOS infants with CONS was 15 days (median 12 days; range three to 54 days), the mean birth weight (+/- SD) was 904+/-247 g, and the mean gestational age at birth (+/- SD) was 27+/-2 weeks. All infants were started on cloxacillin and gentamicin, and improved clinically over the first 48 h. Six isolates were sensitive to cloxacillin. Twenty-three infants grew oxacillin-resistant CONS, eight of whom had persistence of CONS on repeat culture secondary to central lines. Two infants grew two strains of CONS. Eighteen of 22 infants (82%) with in vitro oxacillin-resistant CONS had clearance of bacteremia with cloxacillin and gentamicin. Ten infants (37%) received vancomycin, based on the authors' guidelines. There were no cases of prolonged bacteremia requiring rifampicin. Three infants died, but none of the deaths could be attributed to CONS. The authors describe clinical improvement with clearance of CONS using cloxacillin and gentamicin, despite laboratory results suggesting oxacillin resistance. The authors' unit policy was based on clinical response and permitted the continuation of cloxacillin, provided that a repeat blood culture was negative. Vancomycin use was suggested for clinical deterioration or persistence of CONS. These results question the in vitro tests of resistance. Clearance of oxacillin-resistant CONS from the blood points to in vivo sensitivity, while the laboratory testing suggests in vitro resistance. The absence of subsequent positive blood cultures for CONS confirms clearance of this organism. It was demonstrated that cloxacillin (150 mg/kg/day dose), along with gentamicin, can clear CONS from the blood within 48 h. The relationship between in vivo and in vitro sensitivities also needs to be further studied both in the laboratory and in a prospective trial.


[Pelvic exenteration for late complications of radiation-induced pelvic injury: a preliminary study].
Objective: To investigate the safety and efficacy of total pelvic exenteration (TPE) for treating late complications of radiation-induced pelvic injury. Methods: This was a descriptive case series study. The inclusion criteria were as follows: (1) confirmed radiation-induced pelvic injury after radiotherapy for pelvic malignancies; (2) late complications of radiation-induced pelvic injury, such as bleeding, perforation, fistula, and obstruction, involving multiple pelvic organs; (3) TPE recommended by a multidisciplinary team; (4) patient in good preoperative condition and considered fit enough to tolerate TPE; and (5) patient extremely willing to undergo the procedure and accept the associated risks. The exclusion criteria were as follows: (1) preoperative or intraoperative diagnosis of tumor recurrence or metastasis; (2) had only undergone diversion or bypass surgery after laparoscopic exploration; and (3) incomplete medical records. Clinical and follow-up data of patients who had undergone TPE for late complications of radiation-induced pelvic injury between March 2020 and September 2022 at the Sixth Affiliated Hospital of Sun Yat-sen University were analyzed. Perioperative recovery, postoperative complications, perioperative deaths, and quality of life 1 year postoperatively were recorded. Results: The study cohort comprised 14 women, nine of whom had recto-vagino-vesical fistulas, two vesicovaginal fistulas, one ileo-vesical fistula and rectal necrosis, one ileo-vesical and rectovaginal fistulas, and one rectal ulcer and bilateral ureteral stenosis. The mean duration of surgery was 592.1±167.6 minutes and the median blood loss 550 (100-6000) mL. Ten patients underwent intestinal reconstruction, and four the Hartmann procedure. Ten patients underwent urinary reconstruction using Bricker's procedure and 7 underwent pelvic floor reconstruction. The mean postoperative hospital stay was 23.6±14.9 days. Seven patients (7/14) had serious postoperative complications (Clavien-Dindo IIIa to IVb), including surgical site infections in eight, abdominopelvic abscesses in five, pulmonary infections in five, intestinal obstruction in four, and urinary leakage in two. Empty pelvis syndrome (EPS) was diagnosed in five patients, none of whom had undergone pelvic floor reconstruction. Five of the seven patients who had not undergone pelvic floor reconstruction developed EPS, compared with none of those who had undergone pelvic floor reconstruction. One patient with EPS underwent reoperation because of a pelvic abscess, pelvic hemorrhage, and intestinal obstruction. There were no perioperative deaths. During 18.9±10.1 months of follow-up, three patients died, two of renal failure, which was a preoperative comorbidity, and one of COVID-19. The remaining patients had gradual and significant relief of symptoms during follow-up. QLQ-C30 assessment of postoperative quality of life showed gradual improvement in all functional domains and general health at 1, 3, and 6 months postoperatively (all P<0.05). Conclusions: TPE is a feasible procedure for treating late complications of radiation-induced pelvic injury combined with complex pelvic fistulas. TPE is effective in alleviating symptoms and improving quality of life. However, the indications for this procedure should be strictly controlled and the surgery carried out only by experienced surgeons.


Vitamin D status and incidence of tuberculosis among contacts of pulmonary tuberculosis patients.
A prospective cohort study was conducted from 2009 to 2012 to assess the relationship between serum baseline 25-hydroxivytamin D (vitamin D) status and the incidence of tuberculosis (TB) among 572 contacts of 89 pulmonary TB patients in Castellon, Spain. Three new cases of pulmonary TB occurred, with an incidence density of 3.6 per 1000 person-years. Mean vitamin D status was 13.7 ng/ml for cases and 25.7 ng/ml for non-cases. Vitamin D status showed a significant inverse association with TB incidence (adjusted HR 0.88, 95%CI 0.80-0.97). This result is in line with the hypothesis that vitamin D deficiency is associated with TB incidence.


Human sperm chromosome complements after microinjection of hamster eggs.
A technique was developed for microinjection of human spermatozoa into golden hamster (Mesocricetus auratus) eggs to obtain human pronuclear chromosome complements. Before microinjection the spermatozoa were treated by brief sonication or incubation in TEST-yolk buffer to reduce motility. Very few sperm chromosome complements developed after sperm treatment with sonication and the frequency of spermatozoa with structural chromosomal abnormalities was exceedingly high (91%). The majority of sperm chromosome complements analysed had multiple breaks and rearrangements. Sperm incubation in TEST-yolk buffer before microinjection provided more analysable sperm karyotypes with a significantly lower frequency of structural chromosomal abnormalities (39%, P less than 0.001). Our results therefore suggest that sonication induces structural chromosomal abnormalities in spermatozoa. Since the frequency of chromosomal abnormalities after microinjection was higher than after sperm fertilization of hamster eggs, it appears that microinjection per se may also increase the frequency of chromosomal abnormalities in spermatozoa. These results are based on small numbers and must be confirmed on larger sample sizes, but our study suggests that microinjection of spermatozoa into eggs should not be recommended for clinical use until fully evaluated.


Global diversity of policy, coverage, and demand of COVID-19 vaccines: a descriptive study.
Hundreds of millions of doses of coronavirus disease 2019 (COVID-19) vaccines have been administered globally, but progress on vaccination varies considerably between countries. We aimed to provide an overall picture of COVID-19 vaccination campaigns, including policy, coverage, and demand of COVID-19 vaccines. We conducted a descriptive study of vaccination policy and doses administered data obtained from multiple public sources as of 8 February 2022. We used these data to develop coverage indicators and explore associations of vaccine coverage with socioeconomic and healthcare-related factors. We estimated vaccine demand as numbers of doses required to complete vaccination of countries' target populations according to their national immunization program policies. Messenger RNA and adenovirus vectored vaccines were the most commonly used COVID-19 vaccines in high-income countries, while adenovirus vectored vaccines were the most widely used vaccines worldwide (180 countries). One hundred ninety-two countries have authorized vaccines for the general public, with 40.1% (77/192) targeting individuals over 12 years and 32.3% (62/192) targeting those ≥ 5 years. Forty-eight and 151 countries have started additional-dose and booster-dose vaccination programs, respectively. Globally, there have been 162.1 doses administered per 100 individuals in target populations, with marked inter-region and inter-country heterogeneity. Completed vaccination series coverage ranged from 0.1% to more than 95.0% of country target populations, and numbers of doses administered per 100 individuals in target populations ranged from 0.2 to 308.6. Doses administered per 100 individuals in whole populations correlated with healthcare access and quality index (R2 = 0.59), socio-demographic index (R2 = 0.52), and gross domestic product per capita (R2 = 0.61). At least 6.4 billion doses will be required to complete interim vaccination programs-3.3 billion for primary immunization and 3.1 billion for additional/booster programs. Globally, 0.53 and 0.74 doses per individual in target populations are needed for primary immunization and additional/booster dose programs, respectively. There is wide country-level disparity and inequity in COVID-19 vaccines rollout, suggesting large gaps in immunity, especially in low-income countries.


Epidemiological Clinical Features and Evolution of Gastroduodenal Ulcer Bleeding in a Tertiary Care Hospital in Spain, during the Last Seven Years.
Background. Gastroduodenal ulcer bleeding is a common medical emergency. The aim of this study was to analyze the characteristics of bleeding episodes and to identify changes in the clinical trends over seven years. Methods. Retrospective observational clinical study on a cohort of 272 consecutive adult patients with peptic ulcer bleeding, during the 2006-2012 period. Results. Mean annual admission rate was 12.8 per 100.000 inhabitants. Men were predominant (71%), with a mean age of 66.6 years. Comorbidities were present in 131 cases (48.2%) and 156 patients (57.4%) had received ulcerogenic drugs. Duodenal ulcer was the commonest location (61%). Endoscopic therapy was necessary in 183 cases (67.3%) and rebleeding occurred in 30 patients (11%). Overall mortality rate was 5.5%, with a significant association with the presence of comorbidities (P < 0.01). There were no differences in trends of annual hospitalization, clinical features at presentation, and outcomes during this 7-years period. Conclusions. Annual hospitalization rates and prognosis of peptic ulcer bleeding have remained unchanged in the study period. This may be due to the fact that the effect of improved approach on this condition is probably counteracted by risk factors such as older age, severe comorbidities, and ulcerogenic drugs consumption, which have also remained stable over recent years.


Human peripheral blood T lymphocyte subpopulations isolated on the basis of their affinity for sheep red blood cells differ in angiogenesis-inducing capability.
Human peripheral blood lymphocytes injected intradermally into X-ray immunosuppressed mice were tested for angiogenesis-inducing capacity. Both T and B lymphocytes evoked angiogenesis of the same intensity. The total T cell population was fractionated into three subpopulations on the basis of their different affinities for sheep red blood cells (SRBC). Cells belonging to the subpopulation of T lymphocytes displaying moderate affinity for SRBC induced angiogenesis of the higher intensity, higher than that induced by cells of the total T lymphocyte population. However, lymphocytes both with the highest and with moderate affinity for SRBC, mixed together, evoked angiogenesis no different from that evoked by cells of the total T lymphocyte population, suggesting that inhibitory interactions occur among T cells.


Structures of proteases for ubiqutin and ubiquitin-like modifiers.
Post-translational modifiers can alter the function of proteins in many different ways. The conjugation of ubiquitin (Ub) and ubiqutin-like modifiers (Ubls) to proteins has been shown to be especially crucial in regulating a variety of cellular processes including the cell cycle, growth control, quality control, localization and many more. It is a highly dynamic process and involves a number of enzymes called E1, E2 and E3. Ub and Ubls are removed from the target proteins by deubiquitinating enzymes (DUBs) or Ubl-specific proteases (ULPs), thereby deconjugation can act as an additional level of control over the ubiquitin-conjugation system. In addition, DUBs and ULPs are responsible for activating Ub and Ubls from their inactive corresponding precursor forms. Here we review recent progress in molecular details of these deconjugating enzymes of Ubls.


A light and scanning electron microscopy study of human direct laser metal forming dental implants.
Direct laser metal forming (DLMF) is a procedure in which a high-power laser beam is directed on a metal powder bed and programmed to fuse particles according to a computer-aided design file, thus generating a thin metal layer. With DLMF, it is now possible to fabricate dental implants with a superficial porous surface. The aim of the present study was to evaluate the peri-implant soft tissues around human-retrieved DLMF dental implants. Collagen fibers, in the form of bundles, were oriented perpendicularly to a distance of 100 μm from the surface, where they became parallel, running in several directions. In some portions, only a few collagen fiber bundles appeared to be oriented perpendicularly or obliquely to the plane of the section. Collagen fibers appeared to form a dense chaotic three-dimensional network running in different, more or less parallel directions to the surface. Under scanning electron microscopy, an intimate contact of the fibrous matrix with the implant surface was evident, and some collagen bundles could be seen to bind directly to the metal surface. By changing the surface microtexture, it was possible to change the response of the peri-implant soft tissues.


[Incidence of somatic mutations of the lymphocytes in subjects working in an environment with cancer risk].
A total of 417 individuals were examined by the Strauss-Albertini method, which is useful to detect the lack of hypoxanthin-guanine-phosphoribosil-transferase (HPRT) located on the X-chromosome. The damage of this locus leads to the loss of the sensitivity of the T-lymphocytes to 6-thioguanine (TG) therefore in the presence of this antimetabolite (TG) the cells will respond to the lectin's (phytohaemagglitinine: PHA) growth stimulatory effect. This defect in the cells can cause mutation, making the cells to be resistant against TG-treatment and during their blastogenesis they will be able to incorporate 3 (H)-thymidine. The number of these labeled cells will give the frequency of the mutant (TG-resistant) cells. The results showed a significant elevation in the number of TG-resistant lymphocytes among uranium miners and among industrial workers exposed to different mutagenic or carcinogenic chemicals when compared to the negative controls. The mutation frequency was elevated also among primary lung cancer patients (positive controls) up to the level measured among industrial workers.


Transcriptome of extracellular vesicles released by hepatocytes.
The discovery that the cells communicate through emission of vesicles has opened new opportunities for better understanding of physiological and pathological mechanisms. This discovery also provides a novel source for non-invasive disease biomarker research. Our group has previously reported that hepatocytes release extracellular vesicles with protein content reflecting the cell-type of origin. Here, we show that the extracellular vesicles released by hepatocytes also carry RNA. We report the messenger RNA composition of extracellular vesicles released in two non-tumoral hepatic models: primary culture of rat hepatocytes and a progenitor cell line obtained from a mouse foetal liver. We describe different subpopulations of extracellular vesicles with different densities and protein and RNA content. We also show that the RNA cargo of extracellular vesicles released by primary hepatocytes can be transferred to rat liver stellate-like cells and promote their activation. Finally, we provide in vitro and in vivo evidence that liver-damaging drugs galactosamine, acetaminophen, and diclofenac modify the RNA content of these vesicles. To summarize, we show that the extracellular vesicles secreted by hepatocytes contain various RNAs. These vesicles, likely to be involved in the activation of stellate cells, might become a new source for non-invasive identification of the liver toxicity markers.


Expanding roles for atorvastatin.
Atherosclerosis, especially when manifested as coronary artery disease (CAD), continues to be the number one cause of mortality and morbidity in developed nations and will soon become so in developing countries. Survivors of an acute heart attack have an increased risk of illness and death that is 1.5-15 times greater than in the general population. Sudden death occurs in myocardial infarction (MI) survivors at a rate 4-6 times greater than in the general population. After an initial recognized MI, 25% of male and 38% of female survivors die within 1 year. Within 6 years after a recognized MI, 18% of men and 35% of women will have a second MI, 7% of men and 6% of women will suffer sudden death, and 22% of men and 46% of women will be disabled with heart failure. Aggressive secondary prevention, therefore, is the key to containing and reversing the "malignant" natural history of CAD, since patients with CAD or CAD risk equivalents are already in the "high risk" category according to the Adult Treatment Panel III (ATP III) of the National Cholesterol Education rogram (NCEP). Treatment of dyslipidemia, especially the reduction of low-density lipoprotein (LDL) cholesterol levels to below 100 mg/dl, was recommended by the 2001 NCEP-ATP Guidelines. In 2004, based on the increasing evidence from several major clinical trials between 2001 and 2004, the NCEP-ATP reaffirmed its LDL goal of < 100 mg/dl in patients with CAD or coronary disease risk equivalents (including multiple risk factors), with an optional LDL goal of < 70 mg/dl in very-high-risk patients (including patients with established coronary heart disease plus other highrisk conditions) Findings from major studies, such as the Treating to New Targets (TNT) study, the Scandinavian Simvastatin Survival Study (4S), the Collaborative Atorvastatin Diabetes Study (CARDS), the Stroke Prevention by Aggressive Reduction in Cholesterol Levels (SPARCL) trial and, more recently, the Lipid-Lowering Arm of the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT-LAA), lend support to the idea that greater LDL cholesterol lowering than that achieved with standard doses of statins may be warranted in patients with CAD and metabolic syndrome, CAD and diabetes, CAD and congestive heart failure, and CAD and renal insufficiency. On the other hand, additional lipid reduction may also be warranted in patients with risk factors such as diabetes, hypertension or a history of stroke, but without manifest CAD and despite relatively normal cholesterol levels. These newer indications for statins, atorvastatin in particular, as part of more aggressive secondary and primary prevention, are reviewed in this paper.


Role of poly(ADP-ribose)polymerase in cisplatin-induced injury in LLC-PK1 cells.
Acute renal failure is a dose-limiting factor during cisplatin chemotherapy. We have previously shown in rats that the hydroxyl radical scavenger edaravone reverses cisplatin-induced in vivo renal damage. In the present study, the role of poly(ADP-ribose) polymerase (PARP) in cisplatin nephrotoxicity was investigated in porcine tubular cells LLC-PK1. Cell injury was elicited by transient exposure to 500 microM cisplatin for 1 h or continuous exposure to 30 microM cisplatin for 24 h. Various hydroxyl radical scavengers reversed cell damage in a transient but not permanent model. The cell injury seemed to be necrosis and apoptosis in transient and permanent models, respectively, as assessed by TUNEL method and Annexin V stain. PARP inhibitors such as 3-aminobenzamide and benzamide inhibited cell damage in transient but not permanent model. PARP-dependent cell injury was also observed after transient exposure to hydroxyl radical-generating solution. We demonstrated for the first time the activation of PARP in renal tubular cells by transient cisplatin exposure, as determined by immunofluorescent stain with anti-poly(ADP-ribose) antibody. Moreover, ATP was depleted by transient exposure to cisplatin or hydroxyl radical, both of which were reversed by PARP inhibitors. These findings suggest that hydroxyl radical generation followed by PARP activation contributes to the necrotic cell injury caused by a transient exposure to cisplatin.


Proton-Detected NMR Spectroscopy of Nanodisc-Embedded Membrane Proteins: MAS Solid-State vs Solution-State Methods.
The structural and dynamical characterization of membrane proteins in a lipid bilayer at physiological pH and temperature and free of crystal constraints is crucial for the elucidation of a structure/dynamics-activity relationship. Toward this aim, we explore here the properties of the outer-membrane protein OmpX embedded in lipid bilayer nanodiscs using proton-detected magic angle spinning (MAS) solid-state NMR at 60 and 110 kHz. [1H,15N]-correlation spectra overlay well with the corresponding solution-state NMR spectra. Line widths as well as line intensities in solid and solution both depend critically on the sample temperature and, in particular, on the crossing of the lipid phase transition temperature. MAS (110 kHz) experiments yield well-resolved NMR spectra also for fully protonated OmpX and both below and above the lipid phase transition temperature.


[The re-emergence of the human African trypanosomiasis in Kerou, Benin].
Located in the northern part of Bénin, the district of Kérou is an historical HTA focus of the 60s formerly called the "Atacora focus". This survey was conducted in 2001 to determine the prevalence of HAT in Kérou. The methodology consisted in a cross-sectional survey based on random sampling with two levels of stratification. 3367 persons were included (i=5%). After a screening based on CATT test with total blood test, the examination of trypanosomae was performed with QBC on the subjects that have some persistent antibodies above serum dilution at 1/4, followed by lumbar puncture. For 3367 surveyed subjects, the CATT seroprevalence test with total blood sample is 4.2% and it is 2.4% with serum dilution at 1/8. The research of trypanosomae with QBC is positive in 48 patients and the prevalence is 1.4%. The community survey conducted among 106 positive persons with CATT test serum at 1/4 dilution has revealed that 71 (67%) persons have never left the area since their birth. The HAT is actually emerging in Atacora district in the north of Bénin, especially in Kérou.


Fibrillin assembly requires fibronectin.
Fibrillins constitute the major backbone of multifunctional microfibrils in elastic and nonelastic extracellular matrices. Proper assembly mechanisms are central to the formation and function of these microfibrils, and their properties are often compromised in pathological circumstances such as in Marfan syndrome and in other fibrillinopathies. Here, we have used human dermal fibroblasts to analyze the assembly of fibrillin-1 in dependence of other matrix-forming proteins. siRNA knockdown experiments demonstrated that the assembly of fibrillin-1 is strictly dependent on the presence of extracellular fibronectin fibrils. Immunolabeling performed at the light and electron microscopic level showed colocalization of fibrillin-1 with fibronectin fibrils at the early stages of the assembly process. Protein-binding assays demonstrated interactions of fibronectin with a C-terminal region of fibrillin-1, -2, and -3 and with an N-terminal region of fibrillin-1. The C-terminal half of fibrillin-2 and -3 had propensities to multimerize, as has been previously shown for fibrillin-1. The C-terminal of all three fibrillins interacted strongly with fibronectin as multimers, but not as monomers. Mapping studies revealed that the major binding interaction between fibrillins and fibronectin involves the collagen/gelatin-binding region between domains FNI(6) and FNI(9).


Src-dependent phosphorylation of the epidermal growth factor receptor on tyrosine 845 is required for zinc-induced Ras activation.
Previous studies have shown that exposure of cells to Zn2+ ions induces Ras and MAPK activation through the EGF receptor (EGFR). To further determine the role of EGFR in Zn2+-induced signaling, mouse B82L fibroblasts expressing no detectable EGFR protein (B82L-par), wild type EGFR (B82L-wt), kinase-deficient EGFR (B82L-K721M), or COOH-truncated EGFR (B82L-c'958) were tested. Exposure to Zn2+ induced Ras activity in B82L-wt, B82L-K721M, and B82L-c'958 but not in B82L-par cells, indicating that the tyrosine kinase domain and the auto-phosphorylation sites of the EGFR were not required for Zn2+-induced Ras activation. Zn2+ induced Src activation in all B82L cell lines, including B82L-par, indicating that Src activation is independent of the presence of the EGFR. A Src kinase inhibitor blocked Zn2+-induced Ras activation in all the B82L cell lines capable of this response, suggesting the involvement of Src kinase in Zn2+-induced Ras activation via the EGFR. Zn2+ induced the association of the EGFR with Src and specifically increased the phosphorylation of EGFR at tyrosine 845 (Tyr-845), a known Src phosphorylation site. Stably transfected B82L cells with a point mutation of the EGFR at Tyr-845 (B82L-Y845F) exhibited only basal Ras activity following exposure to Zn2+. These data demonstrate that Src-dependent phosphorylation of the EGFR at Tyr-845 is required for EGFR transactivation and Zn2+-induced Ras activation.


Short DNA sequences from the cytoplasm of mouse tumor cells induce immortalization of human lymphocytes in vitro.
Cytoplasts of mouse L929 and Ehrlich ascites tumor cells harbor DNA sequences that induce unlimited proliferation ("immortalization") of human lymphocytes after transfection in vitro. By equilibrium centrifugation of cytoplasmic lysates in a neutral CsCl gradient, the immortalizing activity was recovered together with extramitochondrial fractions at high salt densities (1.85-1.87 g/cm3). Unexpectedly, these fractions contain linear DNA molecules of 50-500 bp in length. In contrast, cytoplasts of primary, senescent cells (mouse embryo fibroblasts, human lymphocytes) do not harbor DNA in the corresponding fractions. Cytoplasmic DNA isolated from high-density fractions of mouse tumor cells was cloned in subset libraries, and of 45 DNA sequences we identified 2 clones--one from L929 cytoplasts (203 bp) and another one from the cytoplasm of Ehrlich ascites cells (372 bp)--that induce unlimited proliferation of human lymphocytes in vitro. Immortalized lymphoid cells harbor 1-5 copies of transfected DNA integrated into chromosomal DNA, whereas about 100 copies were found as episomal DNA in the cytoplasmic fraction. No immortalization could be induced by transfection of nuclear DNA randomly fragmented to 200-500 bp. Although the cloned DNA sedimented at 1.70 g/cm3, after transient transfection into lymphocytes, these DNA sequences form salt-stable complexes that sediment in fractions at the same high density (1.82-1.88 g/cm3) from which they were originally cloned. The high-density banding of these cytoplasmic DNA sequences may be due to association with RNA and/or with (metallo-) proteins in vivo. Since both cloned DNA sequences with immortalizing activity have stop codons for protein translation in all possible reading frames, immortalization may be induced by insertional inactivation or functional suppression of genes that are needed to be expressed during cellular senescence or programmed cell death.


Evaluation of aggressive surgery for carcinoma of the extrahepatic bile ducts.
The records of 80 consecutive patients with extrahepatic bile duct cancer, 45 women and 35 men, median age 70 years (33-89 years), were reviewed. The histologic diagnoses were adenocarcinoma in 45 patients, 34 cholangiocarcinoma and one squamous cell carcinoma. In 34 patients the tumor was located to the confluence, the right or left hepatic duct, in 16 to the middle and in four to the distal portion of the bile duct. In the remaining 26 patients the tumor comprised more than one of these locations (mixed location). Twenty-seven of the 80 patients (34%) were operated on with resection of the tumor. Among patients 70 years of age and younger the resectability rate was 57%. In nine patients the main surgical procedure was bile duct resection, in 15 patients bile duct resection and liver lobe resection, in 2 patients total pancreatectomy and in one local excision were performed. The resection of the tumor was regarded as radical in 12 patients and palliative in 15. The mortality rate was 11% after resection as compared to 30% in patients with nonresectable tumors. The most common postoperative complication was insufficiency of the anastomosis which occurred in seven patients. Three of these patients required reoperation. The median survival time in patients operated on with radical resection was 20 months, palliative resection 7(1/2) months and in patients with nonresectable tumors 2(1/2) months. The quality of life was estimated according to a special schedule and was found to be improved after resection as compared to nonresection. Patients operated with radical resection spent significantly less of their remaining life at hospital as compared to palliatively resected patients or patients with nonresectable tumors.


Laparoscopic right hepatectomy for living donor.
Many institutions have started or are planning to start a purely laparoscopic right hepatectomy (PLRH) for adult living donor recipients but the experience is relatively very limited. The present review will look at the current status of PLRH and go over some of the technical details important for a safe operation. Necessary elements and different strategies to start a safe and reproducible PLRH program will be discussed. Several publications with a relatively large number of cases have been published in the last few years. The initial results of PLRH seem to be comparable to open donor hepatectomy when performed by surgeons with sufficient expertise laparoscopic and donor hepatectomy. With the ongoing accumulation of experience in laparoscopic liver surgery and living donor liver transplantation, it is most likely that PLRH will be performed more widely than the present time. Institutions should implement a step-by-step approach with proctorship, standardization of surgical procedures and a balanced selection criterion for donors for a safe transition from open method to a PLRH program.


Validation of a new assay for α-synuclein detection in cerebrospinal fluid.
Abnormal α-synuclein aggregation and deposition is the pathological hallmark of Parkinson's disease (PD) and dementia with Lewy bodies (DLB), but is also found in Alzheimer disease (AD). Therefore, there is a gaining interest in α-synuclein in cerebrospinal fluid (CSF) as potential biomarker for these neurodegenerative diseases. To broaden the available choices of α-synuclein measurement in CSF, we developed and validated a new assay for detecting total α-synuclein. This novel ELISA uses commercially available antibodies and is based on electrochemiluminescence technology. The assay protocol is straightforward, with short and simple incubation steps, and requires only small amounts of CSF. We validated this assay for precision, parallelism, dilution linearity, specificity, and spike recovery. We further compared it to the newly validated α-synuclein assay from BioLegend by analyzing a set of 50 CSF samples with both assays. The new assay quantifies α-synuclein in CSF with a lower limit of detection of 36.3 pg/mL and shows no cross-reactivity with human β- and γ-synuclein. Results of dilution linearity, parallelism, spike recovery, and precision classify this assay as well suited for α-synuclein detection in human CSF samples. We present a novel assay based on freely available components to quantify total α-synuclein in CSF as an additional method for α-synuclein as a biomarker in neurodegenerative diseases. The assay convinces with its simple and convenient protocol paired with high sensitivity.


Kilowatt-level cladding light stripper for high-power fiber laser.
We designed and fabricated a high-power cladding light stripper (CLS) by combining a fiber-etched CLS with a cascaded polymer-recoated CLS. The etched fiber reorganizes the numerical aperture (NA) distribution of the cladding light, leading to an increase in the leakage power and a flatter distribution of the leakage proportion in the cascaded polymer-recoated fiber. The index distribution of the cascaded polymer-recoated fiber is carefully designed to ensure an even leakage of cladding light. More stages near the index of 1.451 are included to disperse the heat. The CLS is capable of working consistently under 1187 W of cladding light with an attenuation of 26.59 dB, and the highest local temperature is less than 35°C.


Primary SWL Is an Efficient and Cost-Effective Treatment for Lower Pole Renal Stones Between 10 and 20 mm in Size: A Large Single Center Study.
To assess the clinical features, outcomes, complications, and cost-effectiveness of shockwave lithotripsy (SWL), flexible ureterorenoscopy (FURS), and percutaneous nephrolithotomy (PCNL) in the treatment of lower pole (LP) stones (10-20 mm) in a large tertiary referral center. Consecutive patients treated for solitary LP stones (10-20 mm) between 2008 and 2013 were identified from a prospective database. SWL was used as primary treatment in all cases (following a stone multidisciplinary team assessment), with FURS and PCNL reserved for SWL contraindications, failure, or patient choice. "Success" was defined as stone free and/or clinically insignificant stone fragments (≤3 mm) at 1 and 3 months follow-up. Effect of anatomy on SWL success was determined from using CT images and regression analysis. Average cost per treatment modality (including additional second-line treatments) was calculated for each group using the National Health Service England 2014/15 National Tariff Healthcare Resource Group codes. Two hundred twenty-five patients were included (mean age 54.9; median stone size 12 mm). One hundred ninety-eight (88%), 21 (9.3%), and 6 (2.7%) patients underwent SWL, FURS, and PCNL as primary treatments, respectively, for median stone sizes of 12, 12, and 20 mm. Overall success rates were 82.8%, 76.1%, and 66.7%, respectively (p < 0.05). Sixty-three percent of patients undergoing primary SWL were effectively treated after one session. Anatomical analysis determined infundibulopelvic angle and infundibular length to be significantly different in patients effectively treated with SWL (p = 0.04). The average cost per treatment modality was also significantly lower for SWL (£750) than for FURS (£1261) or PCNL (£2658) (p < 0.01). SWL is both an efficacious and cost-effective primary treatment for patients with solitary LP stones (10-20 mm). The majority of patients can be effectively treated with primary SWL in a dedicated stone center, with the benefits of a short length of stay, low complication, and auxiliary treatment rates. The referral of such patients to high-volume lithotripsy centers with demonstrable outcomes should be given due consideration.


No effect of scalp cooling on survival among women with breast cancer.
Scalp cooling can prevent chemotherapy-induced alopecia in some cancer patients. It is not used in all countries. No data are available regarding its impact, if any, on survival. The aim of this study was to compare overall survival according to whether or not scalp cooling was used during neoadjuvant or adjuvant chemotherapy for non-metastatic breast cancer. We conducted a retrospective cohort study of 1,370 women with non-metastatic invasive breast carcinoma who received chemotherapy in the neoadjuvant or adjuvant setting. A total of 553 women who used scalp cooling came from a tertiary breast cancer clinic in Quebec City (diagnosed between 1998 and 2002) and 817 were treated in other hospitals in the province of Quebec (between 1998 and 2003) where scalp cooling was not routinely available. Overall survival of women who used scalp cooling and those who did not was compared using Cox proportional hazards models. Median follow-up for the scalp-cooled and the non-scalp-cooled groups was 6.3 years and 8.0 years, respectively. Overall mortality was no different (adjusted hazard ratio 0.89, 95 % confidence interval: 0.68-1.17, p = 0.40) among scalp-cooled women, compared to those not getting scalp cooling. Among women getting neoadjuvant or adjuvant chemotherapy for non-metastatic breast cancer, scalp cooling used to prevent chemotherapy-induced alopecia had no negative effect on survival. To our knowledge, this is the first study to compare survival of women who used scalp cooling to that of women who did not.


Pinguisoperla, a new fossil genus of Perlidae (Insecta: Plecoptera) from mid-Cretaceous Burmese amber.
A new fossil stonefly genus and species of the family Perlidae, Pinguisoperla yangzhouensis gen. et sp. nov., is proposed as the second known genus from mid-Cretaceous Burmese amber. The new genus is characterized by its dark coloration and the basally enlarged and curved cerci. Morphological characters of the new genus and species are described, illustrated, and compared with related taxa.


[Screening of key genes and inflammatory signalling pathway involved in the pathogenesis of HLA-B27-associated acute anterior uveitis by gene expression microarray].
To investigate the genes and signalling pathways located upstream of the inflammatory processes in human leukocyte antigen (HLA)-B27-associated acute anterior uveitis by gene expression microarray. Experimental study. HLA-B27-positive and-negative monocytes isolated from human peripheral blood were stimulated with Vibrio cholera lipopolysaccharide (LPS). Gene expression microarrays were used to identify the differentially expressed genes. Differentially expressed (DE) genes were testified by real-time PCR and analyzed by a series of bioinformatics-based techniques such as Gene Ontology, Kyoto Encyclopedia of Genes and Genomes. Gene expression microarray analysis revealed marked differences between HLA-B27-positive acute anterior uveitis (AAU) and HLA-B27-negative healthy control peripheral monocytes in the genes that were upregulated in response to LPS stimulation with 1105 genes and 25 genes respectively. Gene Ontology enrichment and pathway analysis indicated that genes participating in protein transport and folding were essential to the inflammatory process. The LPS receptor-Toll-like receptor (TLR)4 induced TLR signalling pathway and pathway related to Vibrio cholerae infection were located upstream of the network and contribute to the overall response. Among the DE genes, PIK3CA, PIK3CB, AKT3, and MAPK1 might play critical roles in inflammation. Equivalent LPS stimulation induces a different response in HLA-B27-positive peripheral monocytes compared to normal control, suggesting that the TLR pathway is involved in the pathogenesis of HLA-B27-associated AAU.


Regulation of Arabidopsis thaliana 4-coumarate:coenzyme-A ligase-1 expression by artificial zinc finger chimeras.
The use of artificial zinc finger chimeras to manipulate the expression of a gene of interest is a promising approach because zinc finger proteins can be engineered to bind any given DNA sequence in the genome. We have previously shown that a zinc finger chimera with a VP16 activation domain can activate a reporter gene in transgenic Arabidopsis thaliana (Sánchez, J.P., Ullman, C., Moore, M., Choo, Y. and Chua, N.H. (2002) Regulation of gene expression in Arabidopsis thaliana by artificial zinc finger chimeras. Plant Cell Physiol. 43, 1465-1472). Here, we report the use of artificial zinc finger chimeras to specifically regulate the 4-coumarate:coenzyme-A ligase-1 (At4CL1) gene in A. thaliana. At4CL1 is a key enzyme in lignin biosynthesis and the down-regulation of At4CL1 can lead to a decrease in lignin content, which has a significant commercial value for the paper industry. To this end, we designed zinc finger chimeras containing either an activation or a repression domain, which bind specifically to the At4CL1 promoter region. Transgenic lines expressing a zinc finger chimera with the VP16 activation domain showed an increase in At4CL1 expression and enzyme activity. In contrast, transgenic lines expressing a chimera with the KOX (KRAB) repression domain displayed repression of At4CL1 expression and enzyme activity. The activation of At4CL1 expression produced an increase in lignin content, and transgenic plant stems showed ectopic lignin distribution. Repression of the At4CL1 gene resulted in reduced lignin content, and lignin distribution in transgenic stems was severely diminished. Our results confirm and extend previous studies of gene regulation using various artificial zinc finger chimeras in animal and plant systems, and show that this system can be used to up- and down-regulate the expression of an endogenous plant gene such as At4CL1.


Positive-Unlabeled Learning for inferring drug interactions based on heterogeneous attributes.
Investigating and understanding drug-drug interactions (DDIs) is important in improving the effectiveness of clinical care. DDIs can occur when two or more drugs are administered together. Experimentally based DDI detection methods require a large cost and time. Hence, there is a great interest in developing efficient and useful computational methods for inferring potential DDIs. Standard binary classifiers require both positives and negatives for training. In a DDI context, drug pairs that are known to interact can serve as positives for predictive methods. But, the negatives or drug pairs that have been confirmed to have no interaction are scarce. To address this lack of negatives, we introduce a Positive-Unlabeled Learning method for inferring potential DDIs. The proposed method consists of three steps: i) application of Growing Self Organizing Maps to infer negatives from the unlabeled dataset; ii) using a pairwise similarity function to quantify the overlap between individual features of drugs and iii) using support vector machine classifier for inferring DDIs. We obtained 6036 DDIs from DrugBank database. Using the proposed approach, we inferred 589 drug pairs that are likely to not interact with each other; these drug pairs are used as representative data for the negative class in binary classification for DDI prediction. Moreover, we classify the predicted DDIs as Cytochrome P450 (CYP) enzyme-Dependent and CYP-Independent interactions invoking their locations on the Growing Self Organizing Map, due to the particular importance of these enzymes in clinically significant interaction effects. Further, we provide a case study on three predicted CYP-Dependent DDIs to evaluate the clinical relevance of this study. Our proposed approach showed an absolute improvement in F1-score of 14 and 38% in comparison to the method that randomly selects unlabeled data points as likely negatives, depending on the choice of similarity function. We inferred 5300 possible CYP-Dependent DDIs and 592 CYP-Independent DDIs with the highest posterior probabilities. Our discoveries can be used to improve clinical care as well as the research outcomes of drug development.


Favorable outcomes of very elderly patients with critical limb ischemia who undergo distal bypass surgery.
To determine the midterm outcomes of distal bypass surgery for very elderly patients, and to determine the ideal candidates for this procedure. Of 268 consecutive patients (328 limbs) with critical limb ischemia who were treated between 2006 and 2013, 106 (126 limbs) underwent distal bypass and were retrospectively reviewed. Nineteen patients (22 limbs) were aged ≥80 years (very elderly group) and 87 patients (104 limbs) were aged <80 years (control group). The baseline characteristics differed between the 2 groups in terms of regular hemodialysis rate (very elderly group, 4 [21%] vs control group, 60 [69%]; P = .0002) and the Charlson comorbidity index (very elderly group, 3.2 ± 1.7 vs control group, 5.0 ± 2.0; P = .0005). According to the Rutherford category of limb ischemia (4/5/6), the very elderly and control groups were classified as 5/17/0 and 11/87/6, respectively (P = .18). Before the surgery, 17 patients (77%) and 67 patients (64%) were ambulatory in the very elderly and control groups, respectively. At follow-up at 29 ± 22 months, the rates of primary (P = .33) and secondary patency (P = .14), limb salvage (P = .50), survival (P = .26), amputation-free survival (P = .42), major adverse limb event and also perioperative death (P = .11), and major adverse cardiovascular events (P = .36) did not significantly differ between the groups. In multivariate analysis, a history of coronary artery disease (hazard ratio [HR], 2.7; 95% confidence interval [CI], 1.3-5.9; P = .005), preoperative nonambulatory status (HR, 4.2; 95% CI, 2.1-8.1; P < .0001), and serum albumin levels <3 g/dL (HR, 2.7; 95% CI, 1.3-5.4; P = .01) were significantly related to poor amputation-free survival. Thirteen patients (59%) remained ambulatory at the latest follow-up. In 91 patients (110 limbs) with tissue loss, the Society for Vascular Surgery lower extremity threatened limb classification system: risk stratification based on Wound, Ischemia, and foot Infection classification stages 3 and 4 negatively affected complete wound healing, according to multivariate analysis (HR, 0.34; 95% CI, 0.20-0.61; P = .0005). A very elderly age should not preclude a patient from undergoing distal bypass surgery. A history of coronary artery disease, a nonambulatory status, and hypoalbuminemia, along with the Wound, Ischemia, and foot Infection classification for patients with tissue loss, should be carefully considered to obtain the most benefit from distal bypass surgery.


Rocker jaws.
Most adult Polynesian mandibles are of the rocker form. Polynesian crania possess a very open cranial base angle and a large upper facial height. The mandibular growth rotations necessary to maintain occlusion in the presence of this cranial morphology lead to development of an exceptionally closed ramus-body angle, with consequent loss of the antegonial notch and appearance of the rocker form.


ATM and p53 combined analysis predicts survival in glioblastoma multiforme patients: A clinicopathologic study.
Glioblastoma is one of the most malignant cancers, with a distinguishing dismal prognosis: surgery followed by chemo- and radiotherapy represents the current standard of care, and chemo- and radioresistance underlie disease recurrence and short overall survival of patients suffering from this malignancy. ATM is a kinase activated by autophosphorylation upon DNA doublestrand breaks arising from errors during replication, byproducts of metabolism, chemotherapy or ionizing radiations; TP53 is one of the most popular tumor suppressor, with a preeminent role in DNA damage response and repair. To study the effects of the immunohistochemical expression of p-ATM and p53 in glioblastoma patients, 21 cases were retrospectively examined. In normal brain tissue, p-ATM was expressed only in neurons; conversely, in tumors cells, the protein showed a variable cytoplasmic expression (score: +,++,+++), with being completely undetectable in three cases. Statistical analysis revealed that high p-ATM score (++/+++) strongly correlated to shorter survival (P = 0.022). No difference in overall survival was registered between p53 normally expressed (NE) and overexpressed (OE) glioblastoma patients (P = 0.669). Survival analysis performed on the results from combined assessment of the two proteins showed that patients with NE p53 /low pATM score had longer overall survival than the NE p53/ high pATM score counterpart. Cox-regression analysis confirmed this finding (HR = 0.025; CI 95% = 0.002-0.284; P = 0.003). Our study outlined the immunohistochemical expression of p-ATM/p53 in glioblastomas and provided data on their possible prognostic/predictive of response role. A "non-oncogene addiction" to ATM for NEp53 glioblastoma could be postulated, strengthening the rationale for development of ATM inhibiting drugs.


Violent death in the pediatric age group: rural and urban differences.
Violent death (homicide and suicide) in the pediatric age group is a major public health problem. A descriptive study was undertaken to review retrospectively the 1077 pediatric coroner's cases in 11 California counties for differences between urban and rural violent death rates. Pediatric violent death was more prevalent in the urban region than in the rural region (P less than 0.0007). High urban homicide rates accounted for most of this difference. Suicide rates were not significantly different (P = 0.18). Seventy-four percent of the violent deaths were in the 15- to 18-year age group, and most of these deaths were caused by firearms (81%). Blacks had the highest homicide and suicide rates. Child abuse was an important cause of death for young children in the urban area only. Socioeconomic factors, cultural differences, high population density, and the availability of firearms were proposed as factors affecting violent death in the pediatric age group.


The Effects of Shiga Toxin 1, 2 and Their Subunits on Cytokine and Chemokine Expression by Human Macrophage-Like THP-1 Cells.
Infection by Shiga toxin (Stx)-producing enterohemorrhagic Escherichia coli (EHEC) results in severe diarrhea, hemorrhagic colitis, and, occasionally, hemolytic-uremic syndrome (HUS). HUS is associated with an increase in pro-inflammatory cytokines and chemokines, many of which are produced by macrophages in the kidneys, indicating that localized host innate immunity likely plays a role in renal pathogenesis. EHEC serotypes may express one or two classes of serologically defined but structurally and functionally-related Shiga toxins called Stx1 and Stx2. Of these, Stx2 appears to be linked to higher rates of HUS than Stx1. To investigate a possible reason for this, we exposed human macrophage-like THP-1 cells to Stx1 or Stx2 and then used the Luminex multiplex system to assess cytokine/chemokine concentrations in culture supernatant solutions. This analysis revealed that, relative to Stx1, Stx2 significantly caused increased expression of GRO, G-CSF, IL-1β, IL-8 and TNFα in macrophage-like THP-1 cells. This was determined to not be due to a difference in cytotoxicity since both Stx1 and Stx2 displayed similar cytotoxic activities on macrophage-like THP-1 cells. These observations indicate that, in vitro, Stx2 can provoke a greater pro-inflammatory response than Stx1 in macrophages and provides a possible partial explanation for higher rates of HUS in patients infected with EHEC strains expressing Stx2. To begin to determine a mechanism for Shiga toxin-mediated cytokine production, we exposed macrophage-like THP-1 cells to Stx1 or Stx2 A and B subunits. Luminex analysis of cytokines in cell culture supernatant solutions demonstrated that neither subunit alone induced a cytokine response in THP-1 cells.


Treatment Options in Oncology.
Multiple treatment strategies exist for many oncologic problems. In this review, we provide a summary of various reasons for the existence of multiple treatment options in oncology, including factors that concern the treating physician (eg, treatment preferences), environmental factors (eg, financial, regulatory, and scientific aspects), and individual patient-specific factors (eg, medical condition, preferences). We demonstrate the vital role of available treatment options and their origins for clinical decision making and patient communication. These aspects are particularly helpful in the process of shared decision making, which is increasingly favored in situations where there are multiple medically reasonable options.


Speech audiometry findings from HIV+ and HIV- adults in the MACS and WIHS longitudinal cohort studies.
The purpose of this study was to compare various speech audiometry measures between HIV+ and HIV- adults and to further evaluate the association between speech audiometry and HIV disease variables in HIV+ adults only. Three hundred ninety-six adults from the Multicenter AIDS Cohort Study (MACS) and Women's Interagency HIV Study (WIHS) completed speech audiometry testing. There were 262 men, of whom 117 (44.7%) were HIV+, and 134 women, of whom 105 (78.4%) were HIV+. Speech audiometry was conducted as part of the standard clinical audiological evaluation that included otoscopy, tympanometry, and pure-tone air- and bone-conduction thresholds. Specific speech audiometry measures included speech recognition thresholds (SRT) and word recognition scores in quiet presented at 40dB sensation level (SL) in reference to the SRT. SRT data were categorized in 5-dB steps from 0 to 25dB hearing level (HL) with one category as ≥30dB HL while word recognition scores were categorized as <90%, 90-99%, and 100%. A generalized estimating equations model was used to evaluate the association between HIV status and both ordinal outcomes. The SRT distributions across HIV+ and HIV- adults were similar. HIV+ and HIV- adults had a similar percentages of word recognition scores <90%, a lower percentage of HIV- adults had 90-99%, but HIV- adults had a higher percentage of 100%. After adjusting for covariables, HIV+ adults were borderline significantly more likely to have a higher SRT than HIV- adults (odds ratio [OR]=1.45, p=0.06). Among HIV+ adults, HIV-related variables (i.e., CD4+ T-cell counts, HIV viral load, and ever history of clinical AIDS) were not significantly associated with either SRT or word recognition score data. There was, however, a ceiling effect for word recognition scores, probably the result of obtaining this measure in quiet with a relatively high presentation level. A more complex listening task, such as speech-in-noise testing, may be a more clinically informative test to evaluate the effects of HIV on speech communication.


Air contaminants and litter fall decomposition in urban forest areas: The case of São Paulo - SP, Brazil.
Urban forests are usually affected by several types of atmospheric contaminants and by abnormal variations in weather conditions, thus facilitating the biotic homogenization and modification of ecosystem processes, such as nutrient cycling. Peri-urban forests and even natural forests that surround metropolitan areas are also subject to anthropogenic effects generated by cities, which may compromise the dynamics of these ecosystems. Hence, this study advances the hypothesis that the forests located at the margins of the Metropolitan Region of São Paulo (MRSP), Brazil, have high concentrations of atmospheric contaminants leading to adverse effects on litter fall stock. The production, stock and decomposition of litter fall in two forests were quantified. The first, known as Guarapiranga forest, lies closer to the urban area and is located within the MRSP, approximately 20km from the city center. The second, Curucutu forest, is located 70km from the urban center. This forest is situated exactly on the border of the largest continuum of vegetation of the Atlantic Forest. To verify the reach of atmospheric pollutants from the urban area, levels of heavy metals (Cd, Pb, Ni, Cu) adsorbed on the litter fall deposited on the soil surface of the forests were also quantified. The stock of litter fall and the levels of heavy metals were generally higher in the Guarapiranga forest in the samples collected during the lower rainfall season (dry season). Non-metric multidimensional scaling multivariate analysis showed a clear distinction of the sample units related to the concentrations of heavy metals in each forest. A subtle difference between the units related to the dry and rainy seasons in the Curucutu forest was also noted. Multivariate Analysis of Variance revealed that both site and season of the year (dry or rainy) were important to differentiate the quantity of heavy metals in litter fall stock, although the analysis did not show the interaction between these two factors. Precipitation appeared to be an important factor to disperse air pollutants; one method to better regulate this process is the development and integration of green infrastructure at city level, which might contribute to nature-based solutions. Results suggest that although the Curucutu forest is not very far from the MRSP, which could result in heavy metal levels similar to those observed in the Guarapiranga forest, the weather conditions, geographic location and rainfall rates might act as efficient physical barriers against the dispersion of pollutants in the urban area. However, it is important to highlight that in the period studied (2012-2013), MRSP presented unusual features during the winter period marked by the highest levels of precipitation which was due to several numbers of frontal systems and also due to their permanence for a couple days in the region. Thus, it is recommended to continue this study in order to obtain a database for characterizing the seasonal variation of air pollution levels in the litter fall and their adverse effects on ecosystem processes in these remnants of the Atlantic Forest.


Nickel chloride for intracellular staining of neurons in insects.
The advantages of nickel chloride (NiCl2) for intracellular staining of insect neurons are described. Nickel shares with cobalt the features of rapid migration, easy precipitation, and the capability of being silver-intensified. But, nickel-filled electrodes also have lower average resistances and pass current more readily than comparable electrodes filled with cobalt, making it the ion of choice for intracellular marking in many instances.


Forest: Stay Focused app.
Did you know one sixth of smartphone owners check their mobiles 50 times a day? The Forest app is clearly aimed at those types of people.


Cultural Activation of Consumers.
This column discusses "cultural activation," defined as a consumer's recognition of the importance of providing cultural information to providers about cultural affiliations, challenges, views about, and attitudes toward behavioral health and general medical health care, as well as the consumer's confidence in his or her ability to provide this information. An aid to activation, "Cultural Activation Prompts," and a scale that measures a consumer's level of activation, the Cultural Activation Measurement Scale, are described. Suggestions are made about ways to introduce cultural activation as a component of usual care.


Dynamic ventral disc contraction is necessary for Giardia attachment and host pathology.
Giardia lamblia is a common parasitic protist that infects the small intestine and causes giardiasis, resulting in diarrhea, vomiting, weight loss, and malabsorption. Giardiasis leads to cellular damage, including loss of microvilli, disruption of tight junctions, impaired barrier function, enzyme inhibition, malabsorption, and apoptosis. In the host, motile Giardia trophozoites attach to the duodenal microvilli using a unique microtubule organelle called the ventral disc. Despite early observations of disc-shaped depressions in microvilli after parasite detachment, little is known about disc-mediated attachment mechanisms and there little direct evidence showing that parasite attachment causes cellular damage. However, advancements in in vitro organoid models of infection and genetic tools have opened new possibilities for studying molecular mechanisms of attachment and the impact of attachment on the host. Through high-resolution live imaging and a novel disc mutant, we provide direct evidence for disc contraction during attachment, resolving the long-standing controversy of its existence. Specifically, we identify three types of disc movements that characterize contraction, which in combination result in a decrease in disc diameter and volume. Additionally, we investigate the consequences of attachment and disc contractility using an attachment mutant that has abnormal disc architecture. In a human organoid model, we demonstrate that this mutant has a limited ability to break down the epithelial barrier as compared to wild type. Based on this direct evidence, we propose a model of attachment that incorporates disc contraction to generates the forces required for the observed "grasping" of trophozoites on the host epithelium. Overall, this work highlights the importance of disc contractility in establishing and maintaining parasite attachment, leading to intestinal barrier breakdown.


Coronary arterial development: a review of normal and congenitally anomalous patterns.
Coronary artery development is a delicate, complex, and finely tuned process that includes multiple interactions among many pathways, especially in the pericardium and the developing myocardium. There still exists some controversy on the exact origin of certain cellular components. Nevertheless, an understanding of this extremely important developmental process is paramount in identifying some of the causes of anomalous coronary development. There are different patterns of anomalous coronary arteries, with variable risk of myocardial ischemia, malignant arrhythmias, and sudden cardiac death. These anomalies can be broadly categorized into 2 basic anatomic subsets: those with origin of the anomalous coronary artery from the opposite aortic sinus, and those with origin of the anomalous coronary artery from the pulmonary artery. Diagnosis and management of such patterns continues to be challenging. A good knowledge of the normal and abnormal coronary artery development could potentially help us explore new avenues in the treatment of ischemic heart disease as well as anomalous coronary arteries.


Avalanche dynamics in fluid imbibition near the depinning transition.
We study avalanche dynamics and local activity of forced-flow imbibition fronts in disordered media. We focus on the front dynamics as the mean velocity of the interface v is decreased and the pinning state is approached. Scaling arguments allow us to obtain the statistics of avalanche sizes and durations, which become power-law distributed due to the existence of a critical point at v=0 . Results are compared with phase-field numerical simulations.


Veronaea botryosa: molecular identification with amplified fragment length polymorphism (AFLP) and in vitro antifungal susceptibility.
Inter- and intraspecific genomic variability of 18 isolates of Veronaea botryosa originating from clinical and environmental sources was studied using amplified fragment length polymorphism (AFLP). The species was originally described from the environment, but several severe cases of disseminated infection in apparently healthy individuals have been reported worldwide. All tested strains of V. botryosa, identified on the basis of sequencing and phenotypic and physiological criteria prior to our study, were confirmed by AFLP analysis, yielding a clear separation of V. botryosa as a rather homogeneous group from related species. In vitro antifungal susceptibility testing resulted in MIC90s across all strains in increasing order posaconazole (0.25 μg/ml), itraconazole (1 μg/ml), voriconazole (4 μg/ml), terbinafine (4 μg/ml), caspofungin (8 μg/ml), anidulafungin (8 μg/ml), isavuconazole (16 μg/ml), amphotericin B (16 μg/ml), and fluconazole (32 μg/ml). Overall, the isolates showed a uniform pattern of low MICs of itraconazole and posaconazole, but high MICs for remaining agents. The echinocandins (caspofungin and anidulafungin) had no activity against V. botryosa. There was no statistically significant difference between susceptibilities of environmental (n = 11) and clinical (n = 7) isolates of V. botryosa (P > 0.05).


Characterisation of a highly specific, endogenous inhibitor of cysteine protease from Staphylococcus epidermidis, a new member of the staphostatin family.
Staphostatins, a novel family of cysteine protease inhibitors with a unique mechanism of action and distinct protein fold has recently been discovered. In this report we describe the properties of Staphylococcus epidermidis staphostatin A (EcpB), a new member of the family. As for other staphostatins, the recombinant S. epidermidis staphostatin A exerted very narrow inhibitory specificity, limited to cysteine protease from the same species. The closely related proteases from S. aureus cleaved the inhibitor at the reactive site peptide bond and inactivated it. The EcpB homologue, S. aureus staphostatin A (ScpB), was also susceptible to proteolytic cleavage at the same site by non-target cysteine proteases. Conversely, S. aureus staphostatin B (SspC) was resistant to such proteolysis. The difference in the susceptibility of individual inhibitors to proteolytic cleavage at the reactive site suggests subtle variations in the mechanism of interaction with cysteine proteases.


Protein kinase A inhibitors prevent the maintenance of hippocampal long-term potentiation.
The possible involvement of cAMP-dependent protein kinase A (PKA) in mechanisms of long-term potentiation of the Schaffer collateral-commissural input of rat CA1 neurones was investigated using several inhibitors in vitro. If 10 microM H-8, 100 nM KT5720 or 50 microM Rp-cAMPs was applied to the bath before a triple 100 Hz/0.5 s tetanization, post-tetanic and short-term potentiation developed almost normally. However, from about 3 h after tetanization the long-term potentiation (LTP) of the field-EPSP declined with respect to the control in an irreversible manner. These data suggest that besides protein kinase C the synergistic activation of PKA is necessary for the maintenance of LTP.


Comparison of tenofovir and entecavir in the development of acute kidney injury in cirrhotic chronic hepatitis B patients with refractory ascites.
Tenofovir disoproxil fumarate (TDF) and entecavir are effective antiviral medications that are recommended as first-line monotherapies for the treatment of chronic hepatitis B (CHB) infection, including decompensated liver cirrhosis with ascites. Acute kidney injury (AKI) commonly occurs in patients with cirrhosis and ascites. The aim of this study was to compare the development of AKI during TDF and entecavir treatment of CHB patients with cirrhotic refractory ascites. From January 2011 to April 2017, we identified patients who were diagnosed with cirrhosis with refractory ascites and received TDF or entecavir treatments at Kaohsiung Chang Gung Memorial Hospital. AKI was defined as an increase in serum creatinine of more than 0.3 mg/dL or 1.5-fold from baseline. All episodes of AKI were recorded and compared between those who received TDF and entecavir. A total of 111 patients were enrolled in this retrospective study, of which 22 patients were treated with TDF and 89 were treated with entecavir. Patients with AKI episodes had a higher proportion of TDF treatment (P = 0.01), male (P = 0.023), hepatocellular carcinoma (P = 0.007), admission (P = 0.045), and mortality (P = 0.018). Logistic regression analysis illustrated that TDF treatment of patients with comorbidity was an independent risk factor for the development of AKI [odds ratio (OR), 3.756; 95% confidence interval (CI), 1.293-10.912; P = 0.015] and hepatorenal syndrome (OR, 7.651; 95% CI, 1.697-34.508; P = 0.008). TDF treatment is a risk factor for AKI and HRS development in cirrhotic patients with refractory ascites in comparison with entecavir treatment, especially in patients with comorbidity.